Part:BBa_K3905010
T7 / gen3 210-3p-517-5p AND-Gate / Luciferase / Double Terminator
This composite part is used for testing the presence of hsa Micro-RNA 517-5p and 210-3p, using AND-gate logic. Both miRNAs must be present to activate this switch. It has a T7 promoter for expression of functional RNA (Part:BBa_K1614000). This is followed by the code for our toehold switch (Part:BBa_K3905006). This toehold switch can act as a riboregulator to inhibit translation until both micro-RNA 210-3p and 517-5p are present. Both of these microRNAs bind to an anti-miRNA (part BBa_K3905007) which forms a complex that presents both target microRNAs as a trigger to the toehold switch. This triggers the toehold switch to unravel, exposing the RBS and start codon, allowing translation of a protein. This protein should be a reporter protein so that the activity of the toehold switch can be detected and presence of the microRNAs can be confirmed. Our chosen reporter protein is luciferase (Part:BBa_I712019). When Luciferin is added, luciferase will catalyse a reaction that produces light. This light can be detected with a luminometer to affirm a positive test.
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal XbaI site found at 44
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23INCOMPATIBLE WITH RFC[23]Illegal XbaI site found at 44
- 25INCOMPATIBLE WITH RFC[25]Illegal XbaI site found at 44
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 108
Illegal SapI.rc site found at 932
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